Previous methods of preparing ligands, such as nucleotides, for use in affinity chromatography have typically coupled the nucleotide to a solid matrix through the N6 amino group on the purine ring, or via a hydroxyl group of the ribose moiety. However, these ligands are not always effective ligands for affinity chromatography, usually because of steric hindrance or the orientation of the ligand on the solid matrix. The studies of the molecular structures of some nucleotide binding proteins, such as kinases, support these findings.
Recently, in an alternative method, the nucleotide, adenosine triphosphate (ATP), was coupled to an affinity resin indirectly through the gamma-phosphate group of ATP via an aminophenyl moiety. Linking ATP to a resin via an aminophenyl group attached to the gamma-phosphate of ATP has advantages over earlier nucleotide affinity media.
However, a need still exists to develop a still more efficient method for synthesis of nucleotide affinity media that are suitable for use in affinity chromatography and screening methods.